Water stress reduces crop production significantly, and climate change has further aggravated the problem mainly in arid and semi-arid regions. This was the first study on the possible effects of β-sitosterol application in ameliorating the deleterious changes in wheat induced by water stress under field condition and drip irrigation regimes. A field experiment with the split-plot design was conducted, and wheat plants were foliar sprayed with four β-sitosterol (BBS) concentrations (0, 25, 75, and 100mgL-1) and two irrigation regimes [50 and 100% of crop evapotranspiration (ETc)]. Water stress without BBS treatment reduced biological yield, grain yield, harvest index, and photosynthetic efficiency significantly by 28.9%, 42.8%, 19.6%, and 20.5% compared with the well-watered plants, respectively. Proline content increased in water-stressed and BSS-treated plants, owing to a significant role in cellular osmotic adjustment. Application of BSS was effective in reducing the generation of hydrogen peroxide (H2O2) and hence the malondialdehyde content significantly in water-stressed and well-watered wheat plants. Application of BSS up-regulated the activity of antioxidant enzymes (SOD, CAT, POD, and APX) significantly and increased the content of tocopherol, ascorbic acid, and carotene thereby reducing the levels of reactive oxygen species. The increased antioxidant system in BSS treated plants was further supported by the expression level of SOD and dehydrin genes in both water-stressed and well-watered plants. In the present study, the application of BBS at 100mgL-1 was beneficial and can be recommended for improving the growth and yield of the wheat crop under water stress.